Optimization of PCR-based diagnostics for human cytomegalovirus.

Various techniques of DNA template preparation for the PCR-based analysis of human CMV in biological fluids have been compared. Structural polymorphism of a CMV DNA segment (part of the major immediate early gene) in clinical isolates is described; the molecular markers (nucleotide substitutions, deletions, insertions) localized in the analyzed amplicon appear to be suitable for molecular-epidemiological studies. A scheme of spreading of the molecular markers in the population is suggested.

Nonisotopic identification of two point mutations in the CYP21 gene responsible for nonclassic 21-hydroxylase deficiency.

A simple nonradioactive method was developed for identification of the Pro-30-Leu and Val-281-Leu mutant alleles in the CYP21B gene. Not only does this approach improve mutation analysis for patients with the late onset form of 21-hydroxylase deficiency, but it also decreases problems with interference by the CYP21A pseudogene sequence.

Affinity identification of organic anion transporters in brush-border membrane vesicles from rat kidney.

The inhibitory properties of bromoacetyl-p-aminohippuric acid as the affinity probe of the organic anion transport system were studied. Bromoacetylated p-aminohippurate was shown to be able to inhibit irreversibly the p-aminohippurate (PAH) uptake in brush-border membrane vesicles. The inhibition depends on both the time of treatment and the affinity probe concentration.

Rat DNA sequence associated with a complex form of DNA polymerase alpha in nonregenerating liver interacts with a ubiquitous transcription/replication factor Oct-1.

A ubiquitous mammalian transcription factor, Oct-1 (also known as OTF-1, NF-A1, OBP100, or NFIII), stimulates the initiation of replication of adenovirus DNA, and may also be involved in the activation of some chromosomal replication origins. If this is true, binding sites for Oct-1 should be present within regions responsible for the initiation of DNA replication. In this study such a binding site has been identified within a 340bp fragment that was originally isolated from a minor fraction of DNA associated with a complexed form of DNA polymerase alpha from nonregenerating rat liver, and which shows autonomous replication sequence activity in a transient transfection assay.

Effect of injection of RNA isolated from normal and epileptic cortexes of rabbits into functioning mollusc neurons.

Epileptogenic foci were formed in rabbit visual cortex by freezing with liquid nitrogen. RNA isolated from the epileptogenic cortex (RNAepl), or from the frontal lobes (RNAcont) was injected into spontaneously active neurons of the mollusc Planorbarius corneus. The amplitude and duration of the spontaneous action potentials generated following the injection of RNAepl were reproducibly higher than those produced following the introduction of RNAcont.